Part:BBa_K540001:Experience

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Applications of BBa_K540001

XMU-China 2017 tested this part, using the GFP reporter BBa_I13504 and measure the green fluorescence intensity.

It seems that the fluorescence intensity does not have a specific relationship with the concentration of Cobalt. There might be some problems with this promoter.

ZJU-China 2021 tested this part, including ion-specificity of Prcn, response kinetics mediated by cobalt concentration and treating time, and caveats of part measurement.

Since activation by nickel of this part is not measured, we first test whether Prcn could respond specifically to cobalt ions or could respond to other ions as well.

The result shows that Prcn had significantly higher response to cobalt and nickel, than to copper, manganese, and zinc of same concentration, which, consistent with previous reports and measurements, implied that Prcn respond specifically to cobalt and nickel.

Secondly, we tested how the expression of Prcn changed with increasing concentration of cobalt.

The result shows that an optimal inducing concentration lies between 100μM to 200μM. And within 15h, the expression of Prcn would not exhibit a decreasing tendency, while the specific trend depends on the particular concentration of cobalt. Consequently, test work by XMU-China 2017 need higher concentration of cobalt to observe significant fluorescence activity.

Thirdly, we caveat that applying Prcn in E.coli in anaerobic environment will lead to incorrect result.

This figure shows our result when we used 1.5mL-Eppendorf tubes to culture E.coli instead of 15mL-round bottom tubes. It shows that Prcn had very low respond to different concentrations of cobalt, which is misleading.

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